TY - JOUR
T1 - Macrophage esterase
T2 - Identification, purification and properties of a chymotrypsin-like esterase from lung that hydrolyses and transfers nonpolar amino acid esters
AU - Rojas-Espinosa, Oscar
AU - Arce-Paredez, Patricia
AU - Dannenberg, Arthur M.
AU - Kamenetz, Rodger L.
N1 - Funding Information:
Dr Rojas-Espinosa was on leave of absence from the Departamento de Immunologia, Escuela Nacional de Ci6ncias Biol(~gicas del Instituto Polit6cnico Nacional, Mexico 17, DF, Mdxico for most of this study, and was partially supported by a fellowship from the COFAA-SEDICT, Mdxico. We are indebted to Dr J. Fred Woessner, Jr, University of Miami School of Medicine, Biochemistry Department, Miami, Fla. 33152, for critically reviewing this manuscript; to Dr J. Ken McDonald, Ames Research Center, National Aeronautics and Space Administration, Moffett Field, Calif. 94035, for advising us on plastein formation and for supplying us with purified diaminopep-tidase I, Gly-Phe-OMe and Gly-Lys-OMe; and to Drs Hamao Umezawa and Takaaki Aoyagi of the Institute of Microbial Chemistry, Tokyo, Japan, for supplying us with Chymostatin, Pepstatin, Leupeptin and Antipain. We are also indebted to Dr Saimon Gordon of Rockefeller University, Dr Hyun S. Shin of Johns Hopkins University, Dr Henry Z. Movat of Toronto University, Dr Ines Mandl of Columbia University and Dr Woessner for the various assays they performed with our purified esterase. This investigation was supported by Grant No. AI-08876 from the United States-Japan Cooperative Medical Science Program of the National Institute of Allergy and Infectious Diseases, United States Public Health Service; Grant No. HE-14153 from the National Heart and Lung Institute, USPHS, for the Johns Hopkins Specialized Research Center on Lung; and by Contract DADA17-72-C-2187 with the Army Medical Research and Development Command.
PY - 1975/9/22
Y1 - 1975/9/22
N2 - A chymotrypsin-like esterase was purified from beef lung. This lysosomal enzyme, not previously characterized, seemed to be composed of two or more forms with molecular weights of about 52 000. It hydrolysed N-benzoyl-dl-phenylalanine β-naphthol ester at acid and neutral pH; it polymerized l-phenyl-alanine methyl ester(Phe-OMe) at neutral pH; and it transferred the Phe-residue from Phe-OMe to hydroxylamine at neutral pH. Phenylmethanesulfonyl fluoride, an inhibitor of hydrolytic enzymes with serine in their catalytic site, inhibited this enzyme, but pepstatin, the cathepsin D (EC 3.4.4.23) inhibitor, did not. Sulfhydryl reagents were not required for activity. Macrophages, especially pulmonary alveolar macrophages, were a rich source of this esterase, so it is likely that the enzyme purified from lung came from its macrophages. The esterase hydrolysed and transferred monoamino acid esters, especially those of the aromatic type. Cathepsin C, the dipeptidyl peptide hydrolase (EC 3.4.14.1), acted only on dipeptide esters and amides. Pancreatic chymotrypsin acted on both monoamino acid and dipeptide esters. The chymotrypsin-like esterase did not hydrolyse hemoglobin, casein, or plasma albumin. Thus its proteolytic activity, if present, must be limited to specific substrates, as yet unknown.
AB - A chymotrypsin-like esterase was purified from beef lung. This lysosomal enzyme, not previously characterized, seemed to be composed of two or more forms with molecular weights of about 52 000. It hydrolysed N-benzoyl-dl-phenylalanine β-naphthol ester at acid and neutral pH; it polymerized l-phenyl-alanine methyl ester(Phe-OMe) at neutral pH; and it transferred the Phe-residue from Phe-OMe to hydroxylamine at neutral pH. Phenylmethanesulfonyl fluoride, an inhibitor of hydrolytic enzymes with serine in their catalytic site, inhibited this enzyme, but pepstatin, the cathepsin D (EC 3.4.4.23) inhibitor, did not. Sulfhydryl reagents were not required for activity. Macrophages, especially pulmonary alveolar macrophages, were a rich source of this esterase, so it is likely that the enzyme purified from lung came from its macrophages. The esterase hydrolysed and transferred monoamino acid esters, especially those of the aromatic type. Cathepsin C, the dipeptidyl peptide hydrolase (EC 3.4.14.1), acted only on dipeptide esters and amides. Pancreatic chymotrypsin acted on both monoamino acid and dipeptide esters. The chymotrypsin-like esterase did not hydrolyse hemoglobin, casein, or plasma albumin. Thus its proteolytic activity, if present, must be limited to specific substrates, as yet unknown.
UR - http://www.scopus.com/inward/record.url?scp=0016786477&partnerID=8YFLogxK
U2 - 10.1016/0005-2744(75)90019-4
DO - 10.1016/0005-2744(75)90019-4
M3 - Artículo
SN - 0005-2744
VL - 403
SP - 161
EP - 179
JO - BBA - Enzymology
JF - BBA - Enzymology
IS - 1
ER -